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规避害虫对甘蔗破坏性影响的新方法。

Novel approaches to circumvent the devastating effects of pests on sugarcane.

机构信息

Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.

Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada.

出版信息

Sci Rep. 2021 Jun 14;11(1):12428. doi: 10.1038/s41598-021-91985-8.

Abstract

Sugarcane (Saccharum officinarum L.) is a cash crop grown commercially for its higher amounts of sucrose, stored within the mature internodes of the stem. Numerous studies have been done for the resistance development against biotic and abiotic stresses to save the sucrose yields. Quality and yield of sugarcane production is always threatened by the damages of cane borers and weeds. In current study two problems were better addressed through the genetic modification of sugarcane for provision of resistance against insects and weedicide via the expression of two modified cane borer resistant CEMB-Cry1Ac (1.8 kb), CEMB-Cry2A (1.9 kb) and one glyphosate tolerant CEMB-GTGene (1.4 kb) genes, driven by maize Ubiquitin Promoter and nos terminator. Insect Bio-toxicity assays were carried out for the assessment of Cry proteins through mortality percent of shoot borer Chilo infuscatellus at 2nd instar larvae stage. During V, V and V generations young leaves from the transgenic sugarcane plants were collected at plant age of 20, 40, 60, 80 days and fed to the Chilo infuscatellus larvae. Up to 100% mortality of Chilo infuscatellus from 80 days old transgenic plants of V generation indicated that these transgenic plants were highly resistant against shoot borer and the gene expression level is sufficient to provide complete resistance against target pests. Glyphosate spray assay was carried out for complete removal of weeds. In V-generation, 70-76% transgenic sugarcane plants were found tolerant against glyphosate spray (3000 mL/ha) under field conditions. While in V-generation, the replicates of five selected lines 4L/2, 5L/5, 6L/5, L8/4, and L9/6 were found 100% tolerant against 3000 mL/ha glyphosate spray. It is evident from current study that CEMB-GTGene, CEMB-Cry1Ac and CEMB-Cry2A genes expression in sugarcane variety CPF-246 showed an efficient resistance against cane borers (Chilo infuscatellus) and was also highly tolerant against glyphosate spray. The selected transgenic sugarcane lines showed sustainable resistance against cane borer and glyphosate spray can be further exploited at farmer's field level after fulfilling the biosafety requirements to boost the sugarcane production in the country.

摘要

甘蔗(Saccharum officinarum L.)是一种商业种植的经济作物,因其茎内成熟节间的高浓度蔗糖而种植。为了应对生物和非生物胁迫以保护蔗糖产量,已经进行了许多研究来开发抗性。甘蔗的产量和质量总是受到蔗螟和杂草损害的威胁。在本研究中,通过遗传修饰甘蔗,表达两种改良的甘蔗螟虫抗性 CEMB-Cry1Ac(1.8 kb)、CEMB-Cry2A(1.9 kb)和一个草甘膦耐受 CEMB-GTGene(1.4 kb)基因,通过玉米泛素启动子和 nos 终止子驱动,从而解决了两个问题,为甘蔗提供了对昆虫和除草剂的抗性。通过 2 龄幼虫期蔗螟 Chilo infuscatellus 的死亡率,进行了杀虫蛋白的生物毒性测定。在 V、V 和 V 代中,在 20、40、60、80 天的甘蔗植株龄时,从转基因甘蔗植株上采集嫩叶,并喂食给 Chilo infuscatellus 幼虫。在 V 代中,80 天龄的转基因植株对 Chilo infuscatellus 的死亡率高达 100%,表明这些转基因植株对蔗螟具有高度抗性,且基因表达水平足以提供对靶标害虫的完全抗性。进行了草甘膦喷雾试验,以彻底清除杂草。在 V 代中,在田间条件下,发现 70-76%的转基因甘蔗植株对草甘膦喷雾(3000 mL/ha)具有耐受性。而在 V 代中,五个选定品系 4L/2、5L/5、6L/5、L8/4 和 L9/6 的重复试验均对 3000 mL/ha 的草甘膦喷雾具有 100%的耐受性。从本研究可以明显看出,CPF-246 甘蔗品种中 CEMB-GTGene、CEMB-Cry1Ac 和 CEMB-Cry2A 基因的表达对蔗螟(Chilo infuscatellus)表现出有效的抗性,并且对草甘膦喷雾也具有高度的耐受性。选定的转基因甘蔗品系对蔗螟表现出可持续的抗性,在满足生物安全要求后,可以在农民田间水平进一步利用,以提高该国的甘蔗产量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/717b/8203629/57602b9bfc30/41598_2021_91985_Fig1_HTML.jpg

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